Temporal variation of recombinant protein expression in Escherichia coli biofilms analysed at single-cell level

• Tipo: Artigo em Revista Científica Internacional
• Data: 2016

Autores

L. C. Gomes (Autor) FEUP; D. Carvalho (Autor) Outra; R. Briandet (Autor) Outra; F. J. Mergulhão (Autor) FEUP

Revista

Título: Process Biochemistry

Vol. 51 Nº 9

Páginas: 1155-1161

ISSN: 1359-5113

Editora: Elsevier

Indexação

ISI Web of Knowledge - 6 Citações

Scopus - 11 Citações

Projetos Associados

LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

Outras Informações

ID Authenticus: P-00K-VX9

DOI: 10.1016/j.procbio.2016.05.016

Abstract (EN): Bioprocesses based on surface-associated microorganisms are emerging in environmental and industrial areas due to the physiological specificities and heterogeneities of biofilm cells. This study describes a simple and accurate method for evaluating the recombinant protein expression at a single-cell scale during Escherichia coli biofilm development. The model recombinant protein used was enhanced the green fluorescent protein (eGFP), as its intrinsic fluorescence allows us to quantify expression at both population and single-cell levels. The specific cell fluorescence intensity sharply increased during the first 4days of biofilm cultivation. Thereafter, it decreased abruptly reaching a low-level plateau until the end of the experiment. During biofilm development, the population became increasingly heterogeneous with regard to eGFP expression. Three distinct biofilm types were observed over the course of the experiment: one with a homogeneous population (days 3-5), the second with a moderately heterogeneous population (days 6-8) and the third with a strongly heterogeneous population (days 9-11). Observation of E. coli biofilms by confocal laser scanning microscopy revealed marked spatial heterogeneity, with the cells actively producing eGFP restricted to the top layer of the biofilm. The proposed methodology allows a fine analysis of the recombinant protein expression within E. coli biofilms, and it may be used to optimize the processing conditions.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 7